The concentration of cholesterol in laying hens eggs that consumed feed mixture enriched with functional ingredients, laying hens eggs that consumed feed mixture that was not enriched and eggs produced on the family farm was determined. In addition to other functional ingredients, fish oil has been added to the feed mixture for laying hens, which has a beneficial effect on increasing the level of omega-3 fatty acids. In relation to the total oil content 0.3, 0.9 and 1.5% fish oil was added. Prepared egg yolk samples were analyzed by high performance liquid chromatography with UV detection. The highest concentration of cholesterol, in relation to the literature data, was determined in eggs produced on family farm and lower concentration in the eggs of laying hens that consumed feed mixture that was not enriched with functional ingredients. In laying hens that consumed feed mixture enriched with functional ingredients, the lowest concentration of cholesterol was found in laying hens eggs that consumed mixture with 0.3% fish oil and the highest concentration in laying hens that consumed mixture with 1.5% fish oil. Comparing the obtained results, it can be seen that in the eggs of laying hens that consumed feed mixture with the addition of fish oil, the cholesterol concentration is slightly lower compared to the eggs of laying hens that consumed mixture that was not enriched but not significantly. Eggs produced on a family farm have the highest cholesterol levels